title: High-efficiency RNA cloning enables accurate quantification of miRNA expression by deep sequencing.
abstract: Small RNA cloning and sequencing is uniquely positioned as a genome-wide approach to quantify miRNAs with single-nucleotide resolution. However, significant biases introduced by RNA ligation in current protocols lead to inaccurate miRNA quantification by 1000-fold. Here we report an RNA cloning method that achieves over 95% efficiency for both 5′ and 3′ ligations. It achieves accurate quantification of synthetic miRNAs with less than two-fold deviation from the anticipated value and over a dynamic range of four orders of magnitude. Taken together, this high-efficiency RNA cloning method permits accurate genome-wide miRNA profiling from total RNAs.
authors.forename: Zhaojie
authors.initials: Z
authors.lastname: Zhang
authors.forename: Jerome E
authors.initials: JE
authors.lastname: Lee
authors.forename: Kent
authors.initials: K
authors.lastname: Riemondy
authors.forename: Emily M
authors.initials: EM
authors.lastname: Anderson
authors.forename: Rui
authors.initials: R
authors.lastname: Yi
mesh_terms: D003001:Cloning, Molecular; D015536:Down-Regulation; D015870:Gene Expression; D005786:Gene Expression Regulation; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods / Q000592:standards; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D059370:RNA Folding; D034622:RNA Interference
chemical_list: D035683:MicroRNAs